Bacterial Artificial Chromosome Mutagenesis Using Recombineering

Figure 2

Evolution of recombineering as a major modification strategy for BACs. The evolution of homologous recombination over time since the 1980s is depicted in green (conventional homologous recombination) and red boxes (recombineering), respectively, while major applications of recombineering are depicted in orange boxes. Conventional homologous recombination was the main approach for BAC modifications between 1980s and early 1990s ((a)–(c)), alongside early recombineering techniques mediated by E. coli strains carrying activated λ or Rac recombination functions ((d)-(e)). Following the establishment of defined recombineering systems in late 1990’s (f), recombineering emerged as the favourable method over conventional homologous recombination, and technologies for recombineering rapidly expanded in the next few years ((g)-(h)). Recombineering has, thereafter, been adopted for use in a wide range of biological applications ((i)–(m)). See text for details.